LOCUS       sequence-69695-e        4989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid for PCR-based transplant of Saccharomyces 
            cerevisiae GAL10/GAL1 bidirectional promoter in yeast..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  AUTHORS   Horecka J, Chu AM, Davis RW
  TITLE     IpO: plasmids and methods for simplified, PCR-based DNA transplant 
            in yeast.
  JOURNAL   Yeast. 2014 May;31(5):185-93. doi: 10.1002/yea.3006. Epub 2014 Mar 
            20.
  PUBMED    24604451
REFERENCE   2  (bases 1 to 4989)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1002/yea.3006"; 
            journalName: "Yeast"; date: "2014-05"; volume: "31"; issue: "5"; 
            pages: "185-93"
COMMENT     Sequence Label: pJH131
FEATURES             Location/Qualifiers
     source          1..4989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        917..1358
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    917..1034
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     1329..1352
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     promoter        1419..1635
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(2144..2163)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2145..2163)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2173..2190)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2173..2189)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2182..2204)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2330..2785)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2467..2488)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2679..2698
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2812..2916
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2917..2985,2986..3777)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3135..3154)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3948..4536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4437..4456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4690..4707
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4824..4845
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4860..4865,4866..4883,4884..4890)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4898..4914
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4903..4925
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4922..4938
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4922..4938
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4957..4977
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4959..4977
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gagctcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgaa
       61 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
      121 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
      181 agcggtttga agcaggcggc ggaagaagta acaaaggaac ctagaggcct tttgatgtta
      241 gcagaattgt catgcaaggg ctccctagct actggagaat atactaaggg tactgttgac
      301 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
      361 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
      421 gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg tctctacagg atctgacatt
      481 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
      541 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
      601 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
      661 cagttattac ccggccgctc tagaccggtg catgcgctag cttatattga attttcaaaa
      721 attcttactt tttttttgga tggacgcaaa gaagtttaat aatcatatta catggcatta
      781 ccaccatata catatccata tctaatctta cttatatgtt gtggaaatgt aaagagcccc
      841 attatcttag cctaaaaaaa ccttctcttt ggaactttca gtaatacgct taactgctca
      901 ttgctatatt gaagtacgga ttagaagccg ccgagcgggc gacagccctc cgacggaaga
      961 ctctcctccg tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc
     1021 gccgcactgc tccgaacaat aaagattcta caatactagc ttttatggtt atgaagagga
     1081 aaaattggca gtaacctggc cccacaaacc ttcaaattaa cgaatcaaat taacaaccat
     1141 aggatgataa tgcgattagt tttttagcct tatttctggg gtaattaatc agcgaagcga
     1201 tgatttttga tctattaaca gatatataaa tggaaaagct gcataaccac tttaactaat
     1261 actttcaaca ttttcagttt gtattacttc ttattcaaat gtcataaaag tatcaacaaa
     1321 aaattgttaa tatacctcta tactttaacg tcaaggagaa aaaactatac cgcggtgtac
     1381 agacgtcctc gagtgtggtt tcagggtcca taaagctttt caattcatct tttttttttt
     1441 tgttcttttt tttgattccg gtttctttga aatttttttg attcggtaat ctccgagcag
     1501 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtggtgtt
     1561 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacctgcagg
     1621 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
     1681 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     1741 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     1801 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     1861 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaagacag aaaatttgct
     1921 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     1981 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     2041 gcggcggaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     2101 aagggctccc tagctactgg agaatatact ggtacccaat tcgccctata gtgagtcgta
     2161 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     2221 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     2281 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga aattgtaagc
     2341 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     2401 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     2461 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     2521 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     2581 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     2641 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     2701 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     2761 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2821 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2881 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2941 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3001 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3061 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3121 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3181 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3241 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3301 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3361 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3421 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3481 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3541 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3601 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3661 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3721 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3781 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3841 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3901 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3961 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4021 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4081 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4141 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4201 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4261 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4321 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4381 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4441 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4501 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4561 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4621 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4681 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     4741 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     4801 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     4861 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     4921 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     4981 caaaagctg
//
